High-throughput Analysis of Single Hematopoietic Stem Cell Proliferation in Microfluidic Cell Culture Arrays

Overview

Journal Nat Methods

Specialties Biomedical Engineering
Pathology

Date 2011 May 24

PMID 21602799

Citations 114

Authors

Veronique Lecault

Michael VanInsberghe

Sanja Sekulovic

David J H F Knapp

Stefan Wohrer

William Bowden

Francis Viel

Thomas Mclaughlin

Asefeh Jarandehei

Michelle Miller

Didier Falconnet

Adam K White

David G Kent

Michael R Copley

Fariborz Taghipour

Connie J Eaves

R Keith Humphries

James M Piret

Carl L Hansen

Affiliations

Soon will be listed here.

Abstract

Heterogeneity in cell populations poses a major obstacle to understanding complex biological processes. Here we present a microfluidic platform containing thousands of nanoliter-scale chambers suitable for live-cell imaging studies of clonal cultures of nonadherent cells with precise control of the conditions, capabilities for in situ immunostaining and recovery of viable cells. We show that this platform mimics conventional cultures in reproducing the responses of various types of primitive mouse hematopoietic cells with retention of their functional properties, as demonstrated by subsequent in vitro and in vivo (transplantation) assays of recovered cells. The automated medium exchange of this system made it possible to define when Steel factor stimulation is first required by adult hematopoietic stem cells in vitro as the point of exit from quiescence. This technology will offer many new avenues to interrogate otherwise inaccessible mechanisms governing mammalian cell growth and fate decisions.

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